10 results on '"Masato Shibamoto"'
Search Results
2. Generation of Fabry cardiomyopathy model for drug screening using induced pluripotent stem cell-derived cardiomyocytes from a female Fabry patient
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Yuki Kuramoto, Masamichi Ito, Issei Komuro, Katsuki Okada, Hiromasa Tojo, Toshihiro Yamaguchi, Masato Shibamoto, Shigeru Miyagawa, Yoshiki Sawa, Atsuhiko T. Naito, Taku Sakai, Jong-Kook Lee, Yasushi Sakata, Tomoaki Higo, and Akito Nakagawa
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0301 basic medicine ,Patients ,Induced Pluripotent Stem Cells ,Cell ,Drug Evaluation, Preclinical ,Globotriaosylceramide ,Cardiomyopathy ,Clone (cell biology) ,030204 cardiovascular system & hematology ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,X Chromosome Inactivation ,medicine ,Humans ,Myocytes, Cardiac ,Induced pluripotent stem cell ,Molecular Biology ,X chromosome ,business.industry ,Trihexosylceramides ,medicine.disease ,Fabry disease ,030104 developmental biology ,medicine.anatomical_structure ,chemistry ,alpha-Galactosidase ,High-content screening ,Cancer research ,Fabry Disease ,Female ,Cardiomyopathies ,Cardiology and Cardiovascular Medicine ,business - Abstract
Background Fabry disease is an X-linked disease caused by mutations in α-galactosidase A (GLA); these mutations result in the accumulation of its substrates, mainly globotriaosylceramide (Gb3). The accumulation of glycosphingolipids induces pathogenic changes in various organs, including the heart, and Fabry cardiomyopathy is the most frequent cause of death in patients with Fabry disease. Existing therapies to treat Fabry disease have limited efficacy, and new approaches to improve the prognosis of patients with Fabry cardiomyopathy are required. Methods and results We generated induced pluripotent stem cell (iPSC) lines from a female patient and her son. Each iPSC clone from the female patient showed either deficient or normal GLA activity, which could be used as a Fabry disease model or its isogenic control, respectively. Erosion of the inactivated X chromosome developed heterogeneously among clones, and mono-allelic expression of the GLA gene was maintained for a substantial period in a subset of iPSC clones. Gb3 accumulation was observed in iPSC-derived cardiomyocytes (iPS-CMs) from GLA activity-deficient iPSCs by mass-spectrometry and immunofluorescent staining. The expression of ANP was increased, but the cell surface area was decreased in iPS-CMs from the Fabry model, suggesting that cardiomyopathic change is ongoing at the molecular level in Fabry iPS-CMs. We also established an algorithm for selecting proper Gb3 staining that could be used for high-content analysis-based drug screening. Conclusions We generated a Fabry cardiomyopathy model and a drug screening system by using iPS-CMs from a female Fabry patient. Drug screening using our system may help discover new drugs that would improve the prognosis of patients with Fabry cardiomyopathy.
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- 2018
3. Targeted Genome Replacement via Homology-directed Repair in Non-dividing Cardiomyocytes
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Shungo Hikoso, Yasushi Sakata, Takamaru Ishizu, Akito Nakagawa, Yasuaki Kohama, Sachio Morimoto, Yuki Masumura, Tomoaki Higo, Shuichiro Higo, Seiji Takashima, Masato Shibamoto, and Mikio Shiba
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Cardiomyopathy, Dilated ,0301 basic medicine ,Fluorescent Antibody Technique ,Gene Expression ,lcsh:Medicine ,Biology ,Genome ,Article ,Cell Line ,Homology directed repair ,Mice ,03 medical and health sciences ,Transduction (genetics) ,Genome editing ,Genes, Reporter ,Animals ,Humans ,CRISPR ,Myocytes, Cardiac ,lcsh:Science ,Gene ,Gene Editing ,Multidisciplinary ,Cas9 ,Cell Cycle ,lcsh:R ,Recombinational DNA Repair ,Gene targeting ,Flow Cytometry ,Molecular biology ,High-Throughput Screening Assays ,Cell biology ,Disease Models, Animal ,030104 developmental biology ,Genetic Loci ,Gene Targeting ,Mutation ,lcsh:Q ,CRISPR-Cas Systems - Abstract
Although high-throughput sequencing can elucidate the genetic basis of hereditary cardiomyopathy, direct interventions targeting pathological mutations have not been established. Furthermore, it remains uncertain whether homology-directed repair (HDR) is effective in non-dividing cardiomyocytes. Here, we demonstrate that HDR-mediated genome editing using CRISPR/Cas9 is effective in non-dividing cardiomyocytes. Transduction of adeno-associated virus (AAV) containing sgRNA and repair template into cardiomyocytes constitutively expressing Cas9 efficiently introduced a fluorescent protein to the C-terminus of Myl2. Imaging-based sequential evaluation of endogenously tagged protein revealed that HDR occurs in cardiomyocytes, independently of DNA synthesis. We sought to repair a pathological mutation in Tnnt2 in cardiomyocytes of cardiomyopathy model mice. An sgRNA that avoided the mutated exon minimized deleterious effects on Tnnt2 expression, and AAV-mediated HDR achieved precise genome correction at a frequency of ~12.5%. Thus, targeted genome replacement via HDR is effective in non-dividing cardiomyocytes, and represents a potential therapeutic tool for targeting intractable cardiomyopathy.
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- 2017
4. Activation of DNA Damage Response and Cellular Senescence in Cardiac Fibroblasts Limit Cardiac Fibrosis After Myocardial Infarction
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Yuki Masumura, Yuki Kuramoto, Tomoaki Higo, Taku Sakai, Masato Shibamoto, Jong-Kook Lee, Atsuhiko T. Naito, Tomokazu Sumida, Issei Komuro, Akito Nakagawa, Toshihiro Yamaguchi, Masamichi Ito, Shungo Hikoso, Katsuki Okada, Shuichirou Higo, and Yasushi Sakata
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musculoskeletal diseases ,Male ,DNA damage ,Cardiac fibrosis ,Myocardial Infarction ,Cellular senescence ,030204 cardiovascular system & hematology ,03 medical and health sciences ,Mice ,0302 clinical medicine ,medicine ,In Situ Nick-End Labeling ,Animals ,Myocytes, Cardiac ,030212 general & internal medicine ,Myocardial infarction ,Cellular Senescence ,Ventricular Remodeling ,business.industry ,virus diseases ,General Medicine ,Fibroblasts ,medicine.disease ,Flow Cytometry ,Juxtacrine signalling ,Fibrosis ,nervous system diseases ,Blockade ,Mice, Inbred C57BL ,Disease Models, Animal ,Atm gene ,Tissue fibrosis ,Cancer research ,Cardiology and Cardiovascular Medicine ,business ,human activities ,DNA Damage - Abstract
Cardiac fibrosis plays an important role in cardiac remodeling after myocardial infarction (MI). The molecular mechanisms that promote cardiac fibrosis after MI are well studied; however, the mechanisms by which the progression of cardiac fibrosis becomes attenuated after MI remain poorly understood. Recent reports show the role of cellular senescence in limiting tissue fibrosis. In the present study, we tested whether cellular senescence of cardiac fibroblasts (CFs) plays a role in attenuating the progression of cardiac fibrosis after MI. We found that the number of γH2AX-positive CFs increased up to day 7, whereas the number of proliferating CFs peaked at day 4 after MI. Senescent CFs were also observed at day 7, suggesting that attenuation of CF proliferation occurred simultaneously with the activation of the DNA damage response (DDR) system and the appearance of senescent CFs. We next cultured senescent CFs with non-senescent CFs and showed that senescent CFs suppressed proliferation of the surrounding non-senescent CFs in a juxtacrine manner. We also found that the blockade of DDR by Atm gene deletion sustained the proliferation of CFs and exacerbated the cardiac fibrosis at the early stage after MI. Our results indicate the role of DDR activation and cellular senescence in limiting cardiac fibrosis after MI. Regulation of cellular senescence in CFs may become one of the therapeutic strategies for preventing cardiac remodeling after MI.
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- 2019
5. Author Correction: Activation of endothelial β-catenin signaling induces heart failure
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Tomokazu Sumida, Akihito Hashimoto, Ralf H. Adams, Akito Nakagawa, Toru Oka, Masato Shibamoto, Yuki Kuramoto, Mutsuo Harada, Hiroshi Akazawa, Ichiro Shiojima, Issei Komuro, Kazutaka Ueda, Tetsuo Noda, Tomoaki Higo, Jong-Kook Lee, Taku Sakai, Atsuhiko T. Naito, Katsuki Okada, Yasushi Sakata, Seitaro Nomura, and Florian P. Limbourg
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Multidisciplinary ,business.industry ,lcsh:R ,lcsh:Medicine ,medicine.disease ,Text mining ,Heart failure ,ComputingMethodologies_DOCUMENTANDTEXTPROCESSING ,Cancer research ,Medicine ,lcsh:Q ,β catenin signaling ,lcsh:Science ,business - Abstract
A correction to this article has been published and is linked from the HTML and PDF versions of this paper. The error has not been fixed in the paper.
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- 2018
6. Phenotypic Screening Using Patient-Derived Induced Pluripotent Stem Cells Identified Pyr3 as a Candidate Compound for the Treatment of Infantile Hypertrophic Cardiomyopathy
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Jong-Kook Lee, Taku Sakai, Katsuki Okada, Hiroyuki Morita, Masamichi Ito, Akihiro Umezawa, Akito Nakagawa, Ichiro Shiojima, Yoshiki Sawa, Masato Shibamoto, Shigeru Miyagawa, Toshihiro Yamaguchi, Tomoaki Higo, Seitaro Nomura, Tomokazu Sumida, Yasushi Sakata, Atsuhiko T. Naito, Issei Komuro, and Yuki Kuramoto
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0301 basic medicine ,Adult ,Male ,medicine.medical_specialty ,Induced Pluripotent Stem Cells ,macromolecular substances ,030204 cardiovascular system & hematology ,Muscle hypertrophy ,Pathogenesis ,03 medical and health sciences ,0302 clinical medicine ,Transient Receptor Potential Channels ,Internal medicine ,medicine ,Prevalence ,Humans ,Mass Screening ,Myocytes, Cardiac ,cardiovascular diseases ,Induced pluripotent stem cell ,business.industry ,Myocardium ,Noonan Syndrome ,Hypertrophic cardiomyopathy ,Genetic disorder ,General Medicine ,Cardiomyopathy, Hypertrophic ,medicine.disease ,Pathophysiology ,030104 developmental biology ,Phenotype ,Child, Preschool ,Mutation ,cardiovascular system ,Etiology ,Cardiology ,Noonan syndrome ,Calcium ,Cardiology and Cardiovascular Medicine ,business - Abstract
Hypertrophic cardiomyopathy (HCM) is a genetic disorder that is characterized by hypertrophy of the myocardium. Some of the patients are diagnosed for HCM during infancy, and the prognosis of infantile HCM is worse than general HCM. Nevertheless, pathophysiology of infantile HCM is less investigated and remains largely unknown. In the present study, we generated induced pluripotent stem cells (iPSCs) from two patients with infantile HCM: one with Noonan syndrome and the other with idiopathic HCM. We found that iPSC-derived cardiomyocytes (iPSC-CMs) from idiopathic HCM patient were significantly larger and showed higher diastolic intracellular calcium concentration compared with the iPSC-CMs from healthy subject. Unlike iPSC-CMs from the adult/adolescent HCM patient, arrhythmia was not observed as a disease-related phenotype in iPSC-CMs from idiopathic infantile HCM patient. Phenotypic screening revealed that Pyr3, a transient receptor potential channel 3 channel inhibitor, decreased both the cell size and diastolic intracellular calcium concentration in iPSC-CMs from both Noonan syndrome and idiopathic infantile HCM patients, suggesting that the target of Pyr3 may play a role in the pathogenesis of infantile HCM, regardless of the etiology. Further research may unveil the possibility of Pyr3 or its derivatives in the treatment of infantile HCM.
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- 2018
7. DNA single-strand break-induced DNA damage response causes heart failure
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Akihito Hashimoto, Peter J. McKinnon, Hiroshi Akazawa, Atsuhiko T. Naito, Yuki Kuramoto, Toshihiro Yamaguchi, Masamichi Ito, Jong-Kook Lee, Issei Komuro, Masato Shibamoto, Tomokazu Sumida, Ichiro Shiojima, Taku Sakai, Shungo Hikoso, Akito Nakagawa, Tomoaki Higo, Yasushi Sakata, Seitaro Nomura, and Katsuki Okada
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0301 basic medicine ,DNA Repair ,DNA damage ,DNA repair ,Science ,General Physics and Astronomy ,Inflammation ,Ataxia Telangiectasia Mutated Proteins ,030204 cardiovascular system & hematology ,Biology ,General Biochemistry, Genetics and Molecular Biology ,Article ,Pathogenesis ,03 medical and health sciences ,XRCC1 ,Gene Knockout Techniques ,Mice ,0302 clinical medicine ,medicine ,Animals ,Myocytes, Cardiac ,DNA Breaks, Single-Stranded ,DNA Single Strand Break ,Heart Failure ,Multidisciplinary ,NF-kappa B ,General Chemistry ,DNA ,medicine.disease ,Cell biology ,body regions ,stomatognathic diseases ,030104 developmental biology ,X-ray Repair Cross Complementing Protein 1 ,Heart failure ,Cytokines ,medicine.symptom ,DNA Damage - Abstract
The DNA damage response (DDR) plays a pivotal role in maintaining genome integrity. DNA damage and DDR activation are observed in the failing heart, however, the type of DNA damage and its role in the pathogenesis of heart failure remain elusive. Here we show the critical role of DNA single-strand break (SSB) in the pathogenesis of pressure overload-induced heart failure. Accumulation of unrepaired SSB is observed in cardiomyocytes of the failing heart. Unrepaired SSB activates DDR and increases the expression of inflammatory cytokines through NF-κB signalling. Pressure overload-induced heart failure is more severe in the mice lacking XRCC1, an essential protein for SSB repair, which is rescued by blocking DDR activation through genetic deletion of ATM, suggesting the causative role of SSB accumulation and DDR activation in the pathogenesis of heart failure. Prevention of SSB accumulation or persistent DDR activation may become a new therapeutic strategy against heart failure., DNA damage response (DDR) is activated in cardiomyocytes of the failing heart, but the type of DNA damage leading to DDR is unclear. Higo et al. show that in mice heart failure is caused in part by unrepaired DNA single-strand breaks in cardiomyocytes, which activate persistent DDR and trigger an NF-κB-dependent cardiac inflammation.
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- 2017
8. Activation of endothelial β-catenin signaling induces heart failure
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Mutsuo Harada, Akihito Hashimoto, Hiroshi Akazawa, Atsuhiko T. Naito, Yuki Kuramoto, Kazutaka Ueda, Ralf H. Adams, Tetsuo Noda, Jong-Kook Lee, Issei Komuro, Masato Shibamoto, Tomokazu Sumida, Ichiro Shiojima, Taku Sakai, Akito Nakagawa, Toru Oka, Tomoaki Higo, Yasushi Sakata, Seitaro Nomura, Florian P. Limbourg, and Katsuki Okada
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0301 basic medicine ,medicine.medical_specialty ,Angiogenesis ,Neuregulin-1 ,Biology ,Article ,Mice ,03 medical and health sciences ,ErbB ,Internal medicine ,medicine ,Animals ,Author Correction ,Autocrine signalling ,Wnt Signaling Pathway ,beta Catenin ,Heart Failure ,Multidisciplinary ,Wnt signaling pathway ,Endothelial Cells ,LRP6 ,LRP5 ,Survival Analysis ,ErbB Receptors ,Disease Models, Animal ,030104 developmental biology ,Endocrinology ,Catenin ,Cancer research ,Neuregulin - Abstract
Activation of β-catenin-dependent canonical Wnt signaling in endothelial cells plays a key role in angiogenesis during development and ischemic diseases, however, other roles of Wnt/β-catenin signaling in endothelial cells remain poorly understood. Here, we report that sustained activation of β-catenin signaling in endothelial cells causes cardiac dysfunction through suppressing neuregulin-ErbB pathway in the heart. Conditional gain-of-function mutation of β-catenin, which activates Wnt/β-catenin signaling in Bmx-positive arterial endothelial cells (Bmx/CA mice) led to progressive cardiac dysfunction and 100% mortality at 40 weeks after tamoxifen treatment. Electron microscopic analysis revealed dilatation of T-tubules and degeneration of mitochondria in cardiomyocytes of Bmx/CA mice, which are similar to the changes observed in mice with decreased neuregulin-ErbB signaling. Endothelial expression of Nrg1 and cardiac ErbB signaling were suppressed in Bmx/CA mice. The cardiac dysfunction of Bmx/CA mice was ameliorated by administration of recombinant neuregulin protein. These results collectively suggest that sustained activation of Wnt/β-catenin signaling in endothelial cells might be a cause of heart failure through suppressing neuregulin-ErbB signaling and that the Wnt/β-catenin/NRG axis in cardiac endothelial cells might become a therapeutic target for heart failure.
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- 2016
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9. Abstract 15981: DNA Single-strand Break-induced DNA Damage Response Causes Heart Failure
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Masato Shibamoto, Atsuhiko T. Naito, Yasushi Sakata, Shungo Hikoso, Issei Komuro, Jong-Kook Lee, and Tomoaki Higo
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business.industry ,DNA damage ,Cancer ,Inflammation ,medicine.disease ,NFKB1 ,Genomic Stability ,Physiology (medical) ,Heart failure ,medicine ,Cancer research ,medicine.symptom ,Cardiology and Cardiovascular Medicine ,business ,Mitosis ,DNA Single Strand Break - Abstract
Introduction: The DNA damage response (DDR) pathway is activated upon DNA damage. In mitotic cells, the DDR plays essential role in maintaining genomic stability and preventing cancer formation. DNA damage and activation of the DDR are also observed in the post-mitotic cardiomyocytes of patients with end-stage heart failure, however, their roles in the pathogenesis of heart failure remains elusive. Methods and Results: We performed transverse aortic constriction (TAC) operation to produce mice model of pressure-overload induced heart failure. Alkaline- and neutral- comet assay revealed that unrepaired DNA single-strand break (SSB), not double-strand break, is accumulated in cardiomyocytes of the failing heart. Mice with cardiomyocyte-specific deletion of XRCC1, a scaffold protein essential for SSB repair, exhibited more severe heart failure and higher mortality after TAC operation. Knockdown of Xrcc1 using siRNA produced SSB accumulation in cardiomyocytes and SSB accumulation induced persistent DDR through activation of ataxia telangiectasia mutated (ATM) kinase. Activated ATM also induced nuclear translocation of NF-κB and increased the expression of inflammatory cytokines. Activation of DDR, nuclear translocation of NF-κB, and increased expression of inflammatory cytokines were also observed in the failing heart and were enhanced in the heart of cardiomyocyte-specific XRCC1 knockout mice. Conclusions: Unrepaired DNA SSB accumulates in post-mitotic cardiomyocytes and plays a pathogenic role in pressure overload-induced heart failure. Approaches that promote efficient SSB repair or suppress aberrant activation of DDR pathway may become a novel therapeutic strategy against heart failure.
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- 2015
10. Disseminated Intravascular Coagulation with Acutely-Thrombosed Popliteal Aneurysm
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Yasuhiro Ichibori, Hideo Shintani, Yoshitaka Okuhara, Masato Shibamoto, Akira Yoshida, Fumi Sato, Takayuki Yamada, and Shinichi Hatsuoka
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Case Report ,General Medicine - Abstract
Disseminated intravascular coagulation (DIC) is an extremely rare complication of acute thrombosis in popliteal aneurysms and makes it difficult to restore the blood flow with thrombolytic therapy or surgical repair. A 75-year-old man with a history of hypertension presented to the emergency department with complaints of right leg pain and bleeding tendency over a 5-day period. The laboratory findings and multislice computed tomography were suggestive of overt DIC caused by acute thrombosis in the right popliteal aneurysm. Successfully treated with medication, he could discharge without surgical or thrombolytic recanalization of the aneurysm.
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- 2010
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